Concentration-dependent dimerization of staphylokinase variants
with engineered surface charges

J 2026

Concentration-dependent dimerization of staphylokinase variants with engineered surface charges

NEMERGUT, Michal; Monika STULAJTEROVA; Rostislav SKRABANA; Andrej HOVAN; L'ubos AMBRO et al.

Základní údaje

Originální název

Concentration-dependent dimerization of staphylokinase variants with engineered surface charges

Autoři

NEMERGUT, Michal; Monika STULAJTEROVA; Rostislav SKRABANA; Andrej HOVAN; L'ubos AMBRO; Alan STRUNGA; Zbyněk PROKOP; Jiří DAMBORSKÝ; Maria TOMKOVA a Erik SEDLAK

Vydání

Protein Science, MALDEN, WILEY-BLACKWELL, 2026, 0961-8368

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.200 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

colloidal stability; differential scanning calorimetry; dimerization mechanisms; protein aggregation; protein kinetic stability; staphylokinase variants; surface charge engineering

Štítky

14314010, 143180, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 12. 5. 2026 10:55, Mgr. Michaela Hylsová, Ph.D.

Anotace

V originále

Staphylokinase (SAK) is a promising third-generation thrombolytic protein, but its clinical potential is limited by immunogenicity and stability concerns. The conformational and colloidal stabilities of four SAK variants-SAK 42D, SAK STAR, and their non-immunogenic derivatives SAK 42D 3A and SAK STAR 3A-were evaluated using differential scanning calorimetry (DSC), dynamic light scattering (DLS), and aggregation kinetics assays. DSC analyses revealed that thermal denaturation of all variants proceeds via two consecutive irreversible steps, with transition parameters strongly dependent on scan rate and protein concentration. SAK STAR variants exhibited markedly exothermic first transitions and reduced scan rate dependence, suggesting stabilization of intermediate states and suppression of aggregation. In contrast, SAK 42D variants exhibited endothermic or weakly exothermic first transitions and a higher aggregation propensity, correlating with reduced conformational stability and formation of less stable dimers. Colloidal stability tests showed that SAK STAR and SAK STAR 3A remained largely aggregation-resistant, whereas SAK 42D and SAK 42D 3A aggregated rapidly at elevated temperatures (>51 degrees C and >38 degrees C, respectively), following apparent second-order kinetics. DLS confirmed concentration-dependent dimerization in all variants, with SAK 42D 3A displaying pronounced polydispersity and instability. We could rationalize this behavior in the context of engineered surface charges. Our results demonstrate that SAK variant stability is shaped by a complex interplay between primary sequence, dimerization behavior, and aggregation propensity, guiding the design of clinically viable thrombolytic agents and their formulations.

Návaznosti

LM2023049, projekt VaV

Název: Český národní uzel Evropské sítě infrastruktur klinického výzkumu

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CZECRIN - Czech National Node to the European Clinical Research Infrastructure Network

LM2023069, projekt VaV

Název: Výzkumná infrastruktura RECETOX

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná infrastruktura RECETOX

857560, interní kód MU
(Kód CEP: EF17_043/0009632)

Název: CETOCOEN Excellence (Akronym: CETOCOEN Excellence)

Investor: Evropská unie, CETOCOEN Excellence, Spreading excellence and widening participation

Citovat

NEMERGUT, Michal; Monika STULAJTEROVA; Rostislav SKRABANA; Andrej HOVAN; L'ubos AMBRO; Alan STRUNGA; Zbyněk PROKOP; Jiří DAMBORSKÝ; Maria TOMKOVA a Erik SEDLAK. Concentration-dependent dimerization of staphylokinase variants with engineered surface charges. Protein Science. MALDEN: WILEY-BLACKWELL, 2026, roč. 35, č. 3, s. 1-14. ISSN 0961-8368. Dostupné z: https://doi.org/10.1002/pro.70494.

@article{2571808,
   author = {Nemergut, Michal and Stulajterova, Monika and Skrabana, Rostislav and Hovan, Andrej and Ambro, L'ubos and Strunga, Alan and Prokop, Zbyněk and Damborský, Jiří and Tomkova, Maria and Sedlak, Erik},
   article_location = {MALDEN},
   article_number = {3},
   doi = {https://doi.org/10.1002/pro.70494},
   keywords = {colloidal stability; differential scanning calorimetry; dimerization mechanisms; protein aggregation; protein kinetic stability; staphylokinase variants; surface charge engineering},
   language = {eng},
   issn = {0961-8368},
   journal = {Protein Science},
   title = {Concentration-dependent dimerization of staphylokinase variants with engineered surface charges},
   url = {https://onlinelibrary.wiley.com/doi/10.1002/pro.70494},
   volume = {35},
   year = {2026}
}

TY  - JOUR
ID  - 2571808
AU  - Nemergut, Michal - Stulajterova, Monika - Skrabana, Rostislav - Hovan, Andrej - Ambro, L'ubos - Strunga, Alan - Prokop, Zbyněk - Damborský, Jiří - Tomkova, Maria - Sedlak, Erik
PY  - 2026
TI  - Concentration-dependent dimerization of staphylokinase variants with engineered surface charges
JF  - Protein Science
VL  - 35
IS  - 3
SP  - 1-14
EP  - 1-14
PB  - WILEY-BLACKWELL
SN  - 09618368
KW  - colloidal stability
KW  - differential scanning calorimetry
KW  - dimerization mechanisms
KW  - protein aggregation
KW  - protein kinetic stability
KW  - staphylokinase variants
KW  - surface charge engineering
UR  - https://onlinelibrary.wiley.com/doi/10.1002/pro.70494
N2  - Staphylokinase (SAK) is a promising third-generation thrombolytic protein, but its clinical potential is limited by immunogenicity and stability concerns. The conformational and colloidal stabilities of four SAK variants-SAK 42D, SAK STAR, and their non-immunogenic derivatives SAK 42D 3A and SAK STAR 3A-were evaluated using differential scanning calorimetry (DSC), dynamic light scattering (DLS), and aggregation kinetics assays. DSC analyses revealed that thermal denaturation of all variants proceeds via two consecutive irreversible steps, with transition parameters strongly dependent on scan rate and protein concentration. SAK STAR variants exhibited markedly exothermic first transitions and reduced scan rate dependence, suggesting stabilization of intermediate states and suppression of aggregation. In contrast, SAK 42D variants exhibited endothermic or weakly exothermic first transitions and a higher aggregation propensity, correlating with reduced conformational stability and formation of less stable dimers. Colloidal stability tests showed that SAK STAR and SAK STAR 3A remained largely aggregation-resistant, whereas SAK 42D and SAK 42D 3A aggregated rapidly at elevated temperatures (>51 degrees C and >38 degrees C, respectively), following apparent second-order kinetics. DLS confirmed concentration-dependent dimerization in all variants, with SAK 42D 3A displaying pronounced polydispersity and instability. We could rationalize this behavior in the context of engineered surface charges. Our results demonstrate that SAK variant stability is shaped by a complex interplay between primary sequence, dimerization behavior, and aggregation propensity, guiding the design of clinically viable thrombolytic agents and their formulations.
ER  -

NEMERGUT, Michal; Monika STULAJTEROVA; Rostislav SKRABANA; Andrej HOVAN; L'ubos AMBRO; Alan STRUNGA; Zbyněk PROKOP; Jiří DAMBORSKÝ; Maria TOMKOVA a Erik SEDLAK. Concentration-dependent dimerization of staphylokinase variants with engineered surface charges. \textit{Protein Science}. MALDEN: WILEY-BLACKWELL, 2026, roč.~35, č.~3, s.~1-14. ISSN~0961-8368. Dostupné z: https://doi.org/10.1002/pro.70494.

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